ABSTRACT
BACKGROUND: Recent studies have shown that rutin presents a heightened interest due to the plethora of biological activities. The major drawback for this phytocompound is the poor water solubility, a parameter that limits its bioavailability. The study aimed to prepare and assess the inclusion complexes of rutin with ß-cyclodextrin (BCD) and hydroxypropyl-ß-cyclodextrin (HPBCD), followed by the evaluation of the antioxidant, antiproliferative and pro-apoptotic activity against B164A5 murine melanoma cell line. METHODS: Inclusion complexes were prepared by kneading method. Scanning electron microscopy, differential scanning calorimetry and X-ray powder diffraction were used in order to assess their formation. Antioxidant activity was determined by DPPH assay. Antiproliferative and pro-apoptotic effects where tested by MTT, respectively Annexin V-PI assays. Physico-chemical methods proved that incorporation took place. RESULTS: Results show that rutin presents antioxidant activity, and complexation increases this property. After 72 h of incubation at the concentration of 100 µM rutin proved to be an active antiproliferative and pro-apoptotic compound against B164A5 cells. CONCLUSION: Incorporation in BCD and HBCD maintained the activity, thus representing an important finding and open path for in vivo experiments.
Subject(s)
2-Hydroxypropyl-beta-cyclodextrin , Antineoplastic Agents , Antioxidants , Rutin , beta-Cyclodextrins , 2-Hydroxypropyl-beta-cyclodextrin/chemistry , 2-Hydroxypropyl-beta-cyclodextrin/pharmacology , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Antioxidants/chemistry , Antioxidants/pharmacology , Apoptosis/drug effects , Calorimetry, Differential Scanning , Cell Line, Tumor , Cell Proliferation/drug effects , Mice , Powder Diffraction , Rutin/chemistry , Rutin/pharmacology , X-Ray Diffraction , beta-Cyclodextrins/chemistry , beta-Cyclodextrins/pharmacologyABSTRACT
The present study was purported to standardize the high-resolution respirometry technique for the analysis of oxidative phosphorylation (OXPHOS) in saponin-skinned cardiac fibers. Preparation of permeabilized myocardial fibers allows the assessment of respiratory function of the entire population of mitochondria in their normal intracellular position. Adult male rat ventricular bundles were permeabilized with saponin (50 microg/ml) and samples (1-3 mg wet weight) were transferred into the Oxygraph-2k (OROBOROS Instr., Austria) chambers containing air saturated incubation buffer in order to measure Complex I (CI) and II (CII) dependent respiration. The following values (expressed in pmol O2 x s(-1) x mg(-1)) were obtained: CI_LEAK, 67.18 +/- 5.12 (CI dependent basal respiration, after glutamate and malate addition); CI_P, 247.37 +/- 49.90 (CI_OXPHOS state after ADP addition); CI_Pc, 252.036 +/- 53.13 (the intactness of the outer mitochondrial membrane assessed after cytochrome c addition); CI+II_P, 342.90 +/- 62.48 (maximum OXPHOS state obtained after succinate addition by activating convergent electron flow from CI+II into the Q junction of the electron transport system (ETS); CII_P, 302.26 +/- 50.16 (CII dependent OXPHOS state obtained after the addition of a CI inhibitor rotenone, with the subsequent entry of electrons from CII only into the Q junction); ETS capacity, 331.11 +/- 62.39 (uncoupled respiration). The standardized technique will allow the systematic characterization of mitochondrial respiratory dysfunction associated with several cardiac pathologies in both animals and humans.
